Evaluating AGN and Decursin Efficacy in Eμ-myc Transgenic Mice

Mice bearing the Eμ-myc transgene were obtained from the Jackson Laboratory (Bar Harbor, ME, USA). The strain was maintained by breeding hemizygous Eμ-myc transgenic males with wild-type C57BL/6 females. To test the efficacy of AGN and decursin, the mice were administered AGN (200 mg/kg) for eight weeks or decursin (10 mg/kg, ChemFaces) for four weeks. After sacrificing the mice, H&E staining were performed as previously described^{47 (link)}. For the immunohistochemical (IHC) staining of Myc in the spleen tissues, a polyclonal anti-Myc primary antibody (Abcam; ab34072), suitably diluted with a protein diluent (Dako), and a polymer-horseradish peroxidase anti-rabbit (Dako) secondary antibody were used followed by 3,3- diaminobenzidine treatment to visualize the proteins. IHC and H&E-stained samples were examined at 100× magnification (scale bar, 100 μm) with an Olympus CX31 microscope (Olympus Corporation, Tokyo, Japan). The representative images were photographed using a digital photomicrographic camera attachment Moticam 2000 (Motic Co. Ltd., Kowloon, Fujian, China) and Motic Images Plus 2.0 software (Motic Co. Ltd., Kowloon, Fujian, China) and then assembled with PowerPoint software (Microsoft, Redmond, WA, U.S.A).